Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning of a Potentially Strain-Specific DNA Probe of prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method

  • Kook, Joong-Ki (Department of Oral Biochemistry, Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Han, Jin-Ju (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Kim, Hwa-Sook (Department of Oral Biochemistry, College of Dentistry, Chosun University) ;
  • Seong, Jin-Hyo (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Kim, Dong-Kie (Oral Biology Research Institute, College of Dentistry, Chosun University) ;
  • Baek, Dong-Heon (Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University) ;
  • Choe, Son-Jin (Department of Oral Microbiology and Immunology, College of Dentistry, Seoul National University)
  • Published : 2003.04.01
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Abstract

The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.

Keywords

References

  1. J. Kor. Acad. Periodont v.32 Study on development of DNA probe for identification of Prevotella intermedia G8-9K-3 Bak, J. S.;S. H. Kim;D. K. Kim;J. H. Seong;B. O. Kim;M. K. Kim;J. K. Kook
  2. J. Endodontics v.25 Sodium dodecyl sulfate-polyactylamide gel electrophoresis and polymerase chain reaction for differentiation of Prevotella intermedia and Prevotella nigrescens Baumgartner, J. C.;K. S. Bae;T. Xia;J. Whitt;L. L. David https://doi.org/10.1016/S0099-2399(06)81164-7
  3. FEMS Microbiol. Lett. v.138 Prevotella intermedia and Prevotella nigrescens serotypes ribotypes and binding characteristics Dahlen, G. G.;J. R. Johnson;R. Gmur.
  4. Oral Microbiol. Immunol. v.6 Use of randomly cloned DNA fragments for the identification of oral spirochetes DiRienzo, J. M.;S. Cornell;H. Boehringer https://doi.org/10.1111/j.1399-302X.1991.tb00457.x
  5. Int. J. Syst. Bacteriol. v.45 Confirmation of the species Prevotella intermedia and Prevotella nigrescens Frandsen, E. V.;K. Poulasen;M. Kilian https://doi.org/10.1099/00207713-45-3-429
  6. Oral Microbiol. Immunol. v.7 Phenotypic characteristics and DNA relatedness in Prevotella intermedia and similar organisms Fukushima, H.;H. Moroi;J. Inoue;T. Onoe;T. Ezaki;E. Yabuuchi;K. P. Leung;C. B. Walker;W. B. Clark;H. Sagawa https://doi.org/10.1111/j.1399-302X.1992.tb00023.x
  7. J. Periodontol. v.65 Characterization of Prevotella intermedia and Prevotella nigrescens isolates from periodontic and endodontic infections Gharvia, S. E.;M. Haapasalo;H. N. Shah;A. Kotiranta;K. Lounatmaa;M. A. Pearce;D. A. Devine https://doi.org/10.1902/jop.1994.65.1.56
  8. Monoclonal Antibodies Against Bacteria v.I Monoclonal antibodies to characterize the antigenic heterogeneity of Bacteroides intermedius Gmur, R.;C. Wyss;A. J. Macario(ed.);E. Conway de Macario(ed.)
  9. J. Microbiol. Biotechnol. v.12 Molecular identification of vaginal Lactobacillus spp. Isolated from Korean women Kim, E. K.;H. S. Yun;C. E. Chang;T. Lin;S. C. Kim;J. S. So;S. I. Pavlova
  10. J. Periodontol. v.71 A rapid DNA probe method for detection of Porphyromonas ginivalis and Actinobavillus actinomycetemcomitans Komiya, A.;T. Kato;T. Nakagawa;A. Saito;J. Takahashi;S. Yamada;K.Okuda https://doi.org/10.1902/jop.2000.71.5.760
  11. FEMS Microbiol. Lett. v.219 A new method for rapid screening of bacterial species-or subspecies-specific DNA probes Kook, J. K.;M. K. Kim;J. H. Seong;D. K. Kim;B. O. Kim;J. C. Park;K. K. Kim;S. J. Choe;B. M. Min https://doi.org/10.1016/S0378-1097(03)00021-1
  12. Bergey's Manual of Systematic Bacteriology (2th ed.) v.1 Identification of procaryotes Krieg, N. R.
  13. Appl. Environ. Microbiol. v.53 Isolation of intact high-molecular-weight DNA by using guanidine isothiocyanate Lippke, J. A.;M. N. Strzempko;F. F. Raia;S. L. Simon;C. K. French
  14. Oral Microbiol. Immunol. v.11 Distribution and genetic analysis of oral Prevotella intermedia and Prevotella nigrescens Matto, J.;M. Saarela;B. von Troil-Linden;E. Kononen;H. Jousimies-Somer;H. Torkko;S. Alaluusua;S. Asikainen https://doi.org/10.1111/j.1399-302X.1996.tb00342.x
  15. J. Med. Microbiol. v.44 Rapid differentiation of Prevotella intermedia and P. nigrescens by 16S rDNA PCR-RFLP Milsom, S. E.;S. V. Sprague;D. Dymock;A. J. Weightman;W. G. Wade https://doi.org/10.1099/00222615-44-1-41
  16. Infect. Immun. v.56 Serological studies of oral Bacteroides intermedius Nakazawa, F.;J. J. Zambon;H. S. Reynolds;R. J. Genco
  17. J. Microbiol. Biotechnol. v.12 Cloning and characteruzation of the L-lactate dehydrogenase gene (ldhL) from Lactobacillys reuteri ATCC55739 Park, J. Y.;S. Y. Park;S. J. Nam;Y. L. Ha;J. H. Kim
  18. Molecular Cloning: A Laboraty Manual(2nd ed.) Sambrook, J.;E. F. Fritsch;T. Maniatis
  19. Int. J. Syst. Bacteriol. v.42 Biochemical and chemical studies on strains designated Prevotella intermedia and proposal of a new pigmented species, Prevotella nigrescens sp. nov. Shah, H. N.;S. E. Gharbia https://doi.org/10.1099/00207713-42-4-542
  20. Clin. Infect. Dis. v.20(suppl. 2) Detection of putatibe periodontal pathogens in subgingival by 16S ribosomal DNA amplification with the polymerase chain reaction Slots, J.;A. Ashimoto;M. J. Flynn;G. Li;C. Chen
  21. J. Clin. Microbiol. v.37 Improved multiplex PCR using conserved and species-specific 16S rRNA gene primers for simultaneous detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis Trans, S. D.;J. D. Rudney
  22. J. Appl. Bacteriol. v.53 Deoxyribonucleic acid among strains of Bacteroides melaninogenicus and related species van Steenbergen, T. J.;C. A. Vlaanderen;J. de Fraaff https://doi.org/10.1111/j.1365-2672.1982.tb04685.x
  23. J. Periodontol. v.62 Microflora and bacterial DNA restriction enzyme analysis in young adults with periodontitis van Steengergen, T. J.;U. Van der Velden;F. Abbas;J. de Graaff https://doi.org/10.1902/jop.1991.62.4.235